Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Caffeic Acid Phenethyl Ester Induces the Expression of NAG-1 via Activating Transcription Factor 3

ATF3를 통한 caffeic acid phenethyl ester에 의한 NAG-1 유전자의 발현 증가

  • Park, Min-Hee (Department of Biological Sciences, Andong National University) ;
  • Chung, Chungwook (Department of Biological Sciences, Andong National University) ;
  • Lee, Seong Ho (Department of Nutrition & Food Science, University of Maryland, College Park) ;
  • Baek, Seung Joon (College of Veterinary Medicine, Seoul National University) ;
  • Kim, Jong Sik (Department of Biological Sciences, Andong National University)
  • 박민희 (국립안동대학교 생명과학과) ;
  • 정정욱 (국립안동대학교 생명과학과) ;
  • 이성호 (미국 메릴랜드대학교 영양식품과학과) ;
  • 백승준 (서울대학교 수의과대학) ;
  • 김종식 (국립안동대학교 생명과학과)
  • Received : 2017.09.21
  • Accepted : 2017.11.24
  • Published : 2018.01.30
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Abstract

Non-steroidal anti-inflammatory drug-activated gene-1 (NAG-1) is a transforming growth factor beta (TGF-${\beta}$) superfamily gene associated with pro-apoptotic and anti-tumorigenic activities. In the present study, we investigated if caffeic acid phenethyl ester (CAPE) derived from propolis could induce the expression of anti-tumorigenic gene NAG-1. Our results indicate that CAPE significantly induced NAG-1 expression in a time- and concentration-dependent manner in HCT116 cells. We also found that CAPE induced NAG-1 expression in a concentration-dependent manner in another human colorectal cancer cell line, LOVO. In addition, CAPE triggered apoptosis, which was detected with Western blot analysis using poly-(ADP-ribose) polymerase antibody. NAG-1 induction by CAPE was not dependent on transcription factor p53, which was confirmed with Western blot analysis using p53 null HCT116 cells. The luciferase assay results indicated that the new cis-elements candidates were located between -474 and -1,086 of the NAG-1 gene promoter. CAPE dramatically induced activating transcription factor 3 (ATF3) expression, but not cAMP response element-binding protein (CREB), which shares the same binding sites with ATF3. The co-transfection experiment with pCG-ATF3 and pCREB showed that only ATF3 was associated with NAG-1 up-regulation by CAPE, whereas CREB had no effect. In conclusion, the results suggest that CAPE could induce the expression of anti-tumorigenic gene NAG-1 mainly through ATF3.

NAG-1 단백질은 TGF-${\beta}$ superfamily 유전자로서 암세포의 apoptosis를 유도하고 항암 활성과 관련이 있는 것으로 알려져 있다. 본 연구에서는 프로폴리스 유래의 파이토케미칼 CAPE (caffeic acid phenethyl ester)에 의한 항암유전자 NAG-1의 발현과 발현조절에 대해 연구하였다. 인간 대장암 세포주 HCT116에서 CAPE의 처리에 의해 농도의존적, 시간의존적으로 NAG-1의 발현이 증가됨을 확인하였다. 게다가, 다른 대장암 세포주인 LOVO 세포주에서도 농도의존적으로 NAG-1의 발현이 증가됨을 확인하였다. p53-null HCT116세포주를 이용한 실험에서 CAPE에 의한 NAG-1의 발현은 전사조절인자인 p53에 의존하지 않음을 증명하였다. 또한, 3가지 종류의 NAG-1 프로모터 construct를 이용한 실험에서, cis-element 후보가 -474와 -1,086사이에 있음을 증명하였다. CAPE에 의해 전사조절인자인 ATF3와 CREB의 발현이 변화되는 지를 확인한 결과, CREB은 전혀 발현이 증가되지 않는 반면 ATF3는 CAPE 처리에 의해 농도의존적으로 발현이 증가함을 확인하였다. 그리고, pCG-ATF3와 pCREB의 cotransfection 실험에서 CREB은 NAG-1의 발현에 영향을 못 미치는 반면, ATF3의 과대발현에 의해 NAG-1의 발현이 증가됨을 확인하였다. 결론적으로, CAPE에 의한 NAG-1의 발현은 주로 전사조절인자인 ATF3를 경유하여 일어남을 시사한다.

Keywords

References

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